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Transcriptional / functional pathway profiling of DC treated +/- poly I:C +/- kinase inhibitors

transcriptomic dataset

Agilent and Affymetrix data were obtained from an expression analysis of DC treated +/- poly I:C +/- kinase inhibitors. We have previously shown that activation of Src family kinases is absolutely required for cytokine production in DC stimulated by agonists of several Toll like receptors. Inhibition of these kinases by the specific inhibitors PP1/2 uncoupled cytokine production from the up-regulation of co-stimulatory molecules in DC.

We investigated gene expression profiles in DC stimulated with PolyI:C, pretreated or not with PP2, microarray analysis was performed. 41 genes were found to be strongly inhibited by PP2; some cytokines, chemokines and the transcription factor IRF-8 are among these genes. 38 genes were partially inhibited by PP2, and the IL-6 gene belongs to this second group. Other 67 genes were not inhibited at all by PP2. The presence of NF-kB inhibitor in this last group confirms that PP2 doesn’t affect the NF-kB pathway from which IkB transcription depends. Another interesting finding is that among the genes that are not inhibited by PP2 there is also the gene of IL-23. To investigate the role of src kinases on IL-23 production, we tested DC supernatants for the presence of IL-12 and IL-23 cytokines after 24 hours of stimulation with R848 (TLR7/8 agonist) or with Poly I:C (TLR3 agonist). We found that PP2 was able to impair IL-12 production, but it did not affect IL-23 production. On the contrary, the amount of IL-23 was higher in PP2-treated MoDC than in untreated cells, especially upon R848 stimulation.

The results obtained with Agilent arrays were compared with those from the Affymetrix platform.



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